The Pneumocystis Genome Project was conceptualized during the 5th International Workshop on Opportunistic Protists in Lille, France, 1997 and funded by the NIH in 1999. As a community, Pneumocystis investigators decided to use the most common rat Pneumocystis population, Pneumocystis carinii f.sp. carinii karyotype form 1, (now Pneumocystis carinii Frenkel 1999) as the primary focus of the project. The goals of the Project were to sequence its genome with the aid of a physical map constructed from cosmid libraries; construct a partial Expressed Sequence Tag (EST) database from organisms harvested from fulminate infection; and perform limited annotation and functional studies. The community would have access to sequences and reagents generated by the project. All goals of the Project have been met with the exception of completion of the whole genome sequence due to unexpected budgetary constraints and technical difficulties. However, additional resources not listed in the original goals have been generated (e.g. interactive website; ongoing annotation; genefinding). The use of chromosome-specific libraries generated by proprietary nanocloning strategies (Lucigen, Inc.) has overcome problems of cloning bias and poor clonability of certain P. carinii genome regions, encountered during the first few years. Using this strategy, we have made considerable progress in completing the genome; 9.3 Mb of high quality sequence with an expected 14 Mb to be completed by the grant's end. All chromosome libraries will have been constructed. One year of funding is requested to complete the sequencing of the remaining chromosomes and assembly of the genome. Thus, there is but a single aim to this proposal: 1. To finish sequencing the genome of pneumocystis carinii and provide a completed assembly. [unreadable] [unreadable] [unreadable]